Loop-Mediated Isothermal Amplification for Bacterial Gene Detection

Take a master mix containing a strand-displacing DNA polymerase, nucleotides, and a double-stranded DNA-binding dye for loop-mediated isothermal amplification or LAMP.

Add primers that contain a reverse-complementary sequence specific to the target bacterial gene.

Distribute the master mix into the tubes.

Add samples containing bacterial DNA templates to the tubes.

Place the tubes inside a LAMP instrument.

Initial denaturation allows the primer to bind to the target DNA.

As the strand-displacing DNA polymerase starts synthesizing a new strand, it displaces the existing strand, eliminating the need for thermal cycling.

The amplified product forms a looped structure due to the reverse-complementary pairing.

This structure contains multiple start sites for the next cycle, facilitating exponential amplification of target DNA.

The dye binds to the amplified DNA and emits fluorescence.

Monitor the fluorescence in real-time to detect the presence of the target bacterial gene in the samples.

Physically separate the areas used for preparing the LAMP Master Mix and adding the DNA templates.

Clean the work surfaces with isopropanol and a DNA and DNase degrading solution. Then, clean the pipettes and tube